1.Regulative Effect of microRNA let-7c on Myotrophin Gene Expression in Rat’s H9c2 Cardiac Myocytes
Yuyao WANG ; Yuxuan WANG ; Xiang ZHAI ; Ming LIU ; Jun XIE
Chinese Circulation Journal 2016;31(12):1215-1218
Objective: To explore weather microRNA let-7c (miR-let-7c) could regulate myotrophin gene expression in rat’s H9c2 cardiac myocytes with possible mechanisms.
Methods: Recombinant plasmid carrying 3′ untranslated region (3′-UTR) of myotrophin and miRNA precursor of let-7c was co-transfected into Hela cells to construct the luciferase reporter system in order to measure luciferase activity. Rat’s H9c2 cardiac myocytes were cultured. The let-7c gene expression was detected by Taqman probe-based real-time PCR after let-7c miRNA precursor or let-7c miRNA inhibitor transfection respectively; protein expressions of myotrophin and nuclear factor-κB (NF-κB) were examined by Western blot analysis.
Results: Luciferase activity examination indicated that compared with recombinant luciferase gene expression carrier (pMIR-MTPN)+miR precursor negative control group, pMIR-MTPN+miR-let-7c miRNA precursor group showed reduced luciferase activity (59.30±9.90) % vs (98.10±15.10) %. Western blot analysis presented that compared with miR negative control group, miR-let-7c precursor group had decreased protein expressions of myotrophin (0.28±0.05) vs (0.90±0.09) and NF-κB (0.25±0.06) vs (0.75±0.07); in contrast, compared with Negative inhibitor group, miR-let-7c inhibitor group had increased protein expressions of myotrophin (1.14±0.09) vs (0.44±0.09) and NF-κB (1.09±0.05) vs (0.71±0.06), allP<0.05.
Conclusion: miR-let-7c could inhibit myotrophin expression via acting on its 3′-UTR domain and may also inlfuence NF-κB signaling pathway in rat’s H9c2 cardiac myocytes.
2.Quality evaluation of Astmgali radix based on multivariate statistical analysis
Yuyao ZHAI ; Wanli DUAN ; Mingxun HU ; Baolin GUO
International Journal of Traditional Chinese Medicine 2022;44(4):416-421
Objective:To explore the feasibility of using multivariate statistical analysis to evaluate the quality of Astmgali radix from different habitats. Methods:The contents of Astragaloside saponinⅠ, Astragalus saponin Ⅱ, Astragaloside Ⅲ and Astragaloside A were determined by UPLC-ELSD. The components of astragalus saponins from different habitats were analyzed by TOPSIS and cluster thermogram.Results:TOPSIS analysis showed that the comprehensive quality of Astmgali radix samples from Shanxi, Gansu and Inner Mongolia was related(were 0.297 3, 0.346 0, 0.322 5), and the whole quality of S13 and S14 from Shanxi and N5 from Inner Mongolia were higher than others. Cluster thermogram showed that Astmgali radix was grouped into three groups according to region, and the quality difference of Astmgali radix was mainly reflected on Astragaloside saponinⅠand Astragalus saponin Ⅱ. Conclusions:The theory of multivariate statistical analysis is perfect, objective and reliable. It can be used as a reference for comprehensive quality evaluation of Traditional Chinese Medicine (TCM), selection of excellent germplasm resources and traceability of origin of TCM.
3.Analysis of Adverse Drug Reactions/Events of TCM Injections in 18 Third Grade Class A Hospitals from Xi ’an during 2003- 2018
Xiaoai SHOU ; Yu HAO ; Bin HU ; Xiaorong XUE ; Yuyao ZHAI ; Jing WANG ; Yongliang HUANG ; Hui MIN
China Pharmacy 2019;30(19):2696-2701
OBJECTIVE: To analyze the characteristics of adverse drug reactions/events (ADR/ADE) caused by TCM injections in 18 third grade class A hospitals from Xi’an, and to provide reference for rational drug use in clinic. METHODS: Totally 304 ADR/ADE cases of TCM injection reported by 18 third grade class A hospitals of Xi’an were analyzed statistically during 2013-2018 in respect of gender and ages, time distribution of ADR/ADE, distribution of ADR/ADE-inducing drugs, organs/systems involved in ADR/ADE and clinical manifestations, grading of ADR/ADE, outcome and relationship evaluation, drug combination. RESULTS: Totally 6 683 cases of ADR/ADE cases were reported in same period, 304 cases were caused by TCM injection (4.55%). Among 304 cases of ADR/ADE, the number of women (162 cases, 53.29%) was slightly higher than that of men (142 cases, 46.71%). The age of patients was mainly over 40 years old (223 cases, 73.35%). Among them, 118 cases (38.82%) were over 60 years old. ADR/ADE occurred within 1 min to 10 days after administration, especially within 30 min after administration (123 cases, 40.46%). ADR/ADE involved 36 varieties, mainly including agent for promoting blood circulation and dredging, agent for reinforcing and tonifying and agent for clearing away heat and detoxification, and Danhong injection accounted for the highest proportion (43 cases, 14.14%). A total of 352 ADR/ADE case times occurred in 304 patients, mainly lesion of skin and appendents (164 case times, 46.59%), followed by cardiovascular system lesions (54 case times, 15.34%) and systemic lesions (51 case times, 14.49%). The main clinical manifestations were mainly pruritus (117 case times), followed by rash (68 case times) and palpitation (34 case times). Among 304 ADR/ADE reports, 26 cases (8.55%) were severe, 8 cases (2.63%) were new ADR/ADE, 302 cases (99.34%) were cured or improved, 2 cases (0.66%) were unknown, but none of them died. 266 cases (87.50%) were evaluated as likely to be related. 46 patients (15.13%) had combined use of drugs, including 9 cases of combined use of TCM injection and 37 cases of combined use of chemical medicine injection. CONCLUSIONS: TCM injections had a high incidence of ADR/ADE due to the complexity of their components, individual differences and clinical use. Most of them were rapid-onset ADR/ADE within 30 min and mild ADR/ADE commonly seen in lesion of skin and its appendents. The incidence could be reduced by rational clinical use and drug monitoring. For cardiovascular diseases and other basic diseases, attention should be paid to distinguishing their primary diseases from ADR/ADE caused by TCM injections, and more attention should be paid to their individualized drug use.
4.The mechanism of curcumin inhibiting the proliferation of clear cell renal cell carcinoma based on network pharmacology and bioinformatics
Wanli DUAN ; Yuyao ZHAI ; Siyuan PAN ; Haiyan YANG ; Yongyi CHENG ; Yi SUN ; Qian DENG
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(2):278-285
【Objective】 To explore and verify the mechanism of curcumin’s inhibition of the proliferation of renal clear cell carcinoma (RCCC) based on network pharmacology and bioinformatics. 【Methods】 We screened common target genes of RCCC and curcumin from PharmMapper and GeneCards databases. We used TCGA database data analysis to screen out common target genes which not only differentially expressed between RCCC tissue samples and normal tissue samples but also affected prognosis. We also used STRING platform to construct curcumin-RCCC targets interaction network, used R software to perform GO biological process analysis and KEGG pathway enrichment analysis based on the above-mentioned screening target proteins. After curcumin and/or active oxygen inhibitor N-acetyl-L-cysteine (NAC) were incubated in renal cancer 786-O and ACHN cells, CCK8 was used to detect the effects of different concentrations of curcumin on cell proliferation and cell viability. Reactive oxygen detection kit (DCFH-DA) was used to detect the level of intracellular reactive oxygen species, and malondialdehyde (MDA) determination kit (TBA method) to detect intracellular malondialdehyde changes. 【Results】 PharmMapper website and GeneCards database screened out 109 common targets of curcumin and RCCC. TCGA database data analysis screened out 37 differentially expressed genes (DEGs) that might affect the overall survival of patients. The core target proteins of curcumin screened out by protein-protein interaction (PPI) that inhibited the biological behavior of RCCC mainly involved CASP3, EGFR, CHEK1, HSP90AA1, and AR. GO enrichment analysis identified 213 items, mainly including reactive oxygen species metabolic process, response to steroid hormones, fibrinolysis and other biologically active processes. KEGG enrichment analysis identified 24 items, which were mainly related to pyruvate metabolism, glycolysis/gluconeogenesis, FoxO signaling pathway, colorectal cancer, tyrosine metabolism, IL-17 signaling pathway, apoptosis and other signaling pathways. Curcumin reduced the cell viability of 786-O and ACHN in a time- and dose-dependent manner (P<0.05). After curcumin was incubated with kidney cancer cells, the level of reactive oxygen species and MDA increased significantly (P<0.05). The addition of NAC reversed the effect of curcumin on the cell viability of 786-O and ACHN cells (P<0.05). 【Conclusion】 Curcumin may participate in the oxidative stress pathway to inhibit the proliferation of renal cell carcinoma.